Fetal Alcohol Syndrome (FAS) and Fetal Alcohol Spectrum Disorders (FASD) cause severe CMS abnormalities, such as impairments in learning, memory, attention, and executive functions, and pose an enormous financial burden to society. A therapeutic preventative treatment has not been identified. The long-term objective of these studies is to identify a well-tolerated, therapeutic treatment, with widespread actions that has the potential to prevent CNS abnormalities associated with in utero ethanol exposure. The proposed studies will investigate the ability of a-lipoic acid (LA), an antioxidant (AO) and natural compound that is used by most cells, to prevent ethanol-augmented apoptosis (cell death) in developing CNS neurons, and determine whether LA's actions are limited to AO effects or also involve the activation of the important phosphatidyl-inositol 3'kinase pro-survival pathway (PI-3K-> pAkt). Previously, this laboratory showed that a serotonin-1 A receptor agonist protected a subclass of fetal neurons from ethanol-associated apoptosis by activating PI-3K -> pAkt and increasing the expression of the NF-KB dependent pro-survival genes XIAP and Bel-XL. Since LA exerts neuroprotective effects and is used by most CNS cells, it has the potential to protect many CNS neurons. We believe LA mediates its effects by dual actions, i.e., as an AO and by activating PI-3K. Thus, it has the potential to be a potent therapeutic agent. We propose to use an in vitro model to investigate the effects of ethanol and LA on fetal neurons. The specific aims will determine if treatment of fetal rhombencephalic neurons with LA 1) attenuates ethanol associated generation of reactive oxygen species (ROS) and prevents an ethanol-associated reduction in the endogenous AO, GSH; 2) activates the PI-3K prosurvival pathway in ethanol-treated neurons; and 3) subsequently increases the expression of NF-KB dependent genes downstream of PI-3K that encode prosurvival proteins or anti-oxidant enzymes. We will use a fluorescent dye to quantify ROS levels, a colorimetric kit to measure GSH, western blots to detect pAkt, and quantitative real-time RT-PCR to measure gene expression. LAY ABSTRACT: These studies hope to identify a treatment to prevent brain damage in Fetal Alcohol Syndrome. We will investigate whether a therapeutic dose of a natural compound (lipoic acid, LA) that is used by most cells can prevent ethanol from causing developing brain cells to die, and determine whether LA prevents this damage by its antioxidant properties and by activating an important cellular survival pathway. [unreadable] [unreadable] [unreadable]